1, 11-iminoestratrienes



United States Patent M 3,251,833 1,11-IMINOESTRATRIENES Edward Warren Cantrall, New City, Ransom Brown Conrow, Pearl River, and Seymour Bernstein, New City, N.Y., assignors to American Cyanamid Company, Stamford, Conn., a corporation of Maine No Drawing. Filed Apr. 23, 1964, Ser. No. 362,185 18 Claims. (Cl. 260-239.5)

This invention relates to new steroid compounds. More particularly, it relates to 1,l1-iminoestra-1,3,5(l0)- trienes and their preparation.

The novel steroids of this invention can be illustrated by the following formula:

MT Li R. J

wherein R is selected from the group consisting of hydrogen and lower alkyl; R is selected from the group consisting of hydrogen, lower alkyl, lower alkanoyl, mono-nuclear aroyl, and lower alkoxy mono-nuclear aroyl; X is selected from the group consisting of C=O;

' OH OH x \C/ and \C/ H 1owerallryl 011:0111 05011 and acid addition and quaternary ammonium salts thereof.

The present compounds are, in general, crystalline solids relatively insoluble in organic solvents such as lower alkyl alcohols, acetone, ethyl acetate, benzene, toluene, chloroform, ether, petroleum ether and the like.

The present compounds which are described broadly as 1,11-iminoestratrienes substituted in the 17-position can be made from the appropriate 2-azidoestratrienes by heating with hexadecane in the absence of oxygen, and by irradiation with ultraviolet light in benzene, cyclohexane and the like. The l-azido-estratrienes are obtained from the corresponding l-aminoestratrienes described and claimed in our copending application Serial No. 362,161 filed April 23, 1964 by treating the latter with an alkali metal nitrite at a low temperature under acid conditions, and then adding an alkali metal azide.

Using the procedure described hereinafter the following 4-amino-derivatives are among those which may be used as starting materials:

4-amino-3-methoxy-l9-norpregna-1,3,5 -trien-20-one;

4-amino-1 1[3-acetoxy-S-methoxyestra-1,3,5 10) -trien- 17-one;

4-amino-l 7,20; 20,21-bismethylenedioxy-3-methoxy-19- norpregna-1,3,5 10)-trien-1 1 8-01 1 l-acetate;

4-amino-3-methoxyestra-1,3,5 10) -trien-l6u,17fl-diol 16,17-diacetate and 4-amino-3-methoxyestra-1,3,5 10) trien-17B-oll 6-one 17-acetate.

The above compounds are prepared from the corresponding compounds lacking a 4-amino substituent by nitration (nitric acid-acetic acid) and methylation (dimethylsulfate), followed by reduction (sodium hydrosulfite); e.g. estrone 4-nitroestrone 4-nitroestrone methyl ether 4-aminoestrone methyl ether.

The 4-amino steroids immediately above may be transformed into the following l-amino-compounds by methods described hereinafter:

1-amino 3-methoxyestra-1,3,5(10)-trien-17-one; 1-amino-3-methoxy-l 9-norpregna-1,3 ,5 10 -trien-20-one;

3,251,833 Patented May 17, 1966 l-amino-l 1 8-acetoxy-3-rnethoxyestra-1 ,3,5 10) -trien-17- one; 7

1-amino-17, 20; 20,2 1 -bismethylenedioxy-3 -methoxy-19- norpregna-l ,3 ,5 10)-trien-1 1 3-01 1 l-acetate';

1-arnino-3-methoxyestra-1,3,5(10)-trien-16a,17fl-diol 16,17-diacetate;

1-amino-3 -methoxyestra-1,3,5 (10) -trien-1718-ol-16-one 17-acetate.

The l-amino-estratrienes described above on treatment with an alkali metal nitrite under acid conditions at a low temperature yield diazonium salts which upon treatment with alkali metal azides will produce for example l-azido- 3-methoxy-1,3,5(10)-estratrien-l7-one. The latter compound on heating with hexadecane in an oxygen-free atmosphere will produce 1,11-imino-3-rnethoxyestra-1,3,5 (10)-trien-17-one; The -1,l1-imino-compound is also prepared by irradiation with ultraviolet light a benzene, cyclohexane, and the like, solution of the l-azido compound. The hydrogen atom on the 1,1l-imino group may be replaced by a lower alkanoyl group by reaction with a lower alkanoyl chloride or anhydride and by a lower alkyl group by reaction with an alkyl halide -in the presence of an alkali metal hydride.

Among the compounds of the present invention are, for example, I

The 1,1l-imino steroids of the present invention when administered to rats produce a lowering of blood cholesterol and therefore are useful as hypocholesterolemic agents. They also show bactericidal and fungicidal activity. v

The following examples describe in detail the preparation of representative 1,11-iminoestratrienes of the present invention.

EXAMPLE 1 1- (p-nitrophenylazo) -3-meth0xy-4-amin0estra-1,3,5(10 trien-l 7 -one To a stirred solution of 7.0 g. (50.5 m. moles) of pnitroaniline in 50 ml. of glacial acetic acid and ml. of 2.0 N hydrochloric acid m. moles) at -5 C. is added a solution of 3.48 g. (50.1 m. moles) of sodium nitrite in 15 m1. of water below the surface of the liquid, and stirring is continued for 15 minutes. The resulting solution of p-nitrobenzene diazonium chloride is poured into a well-stirred solution of 14.97 g. (50.0 m. moles) of 4-amino-3-methoxyestra-1,3,5(10)-trien-17-one (M.P. 189l93 C.) in 250 ml. of glacial acetic acid and 25 ml. of 5.0 N sodium hydroxide (125 m. moles) at room temperature. The deep red mixture is diluted with one liter of water, allowed to stand overnight then filtered and the product washed thoroughly on the filter with water. The yield of crude material after drying was 21.0 g. (94%), MP. -l90 C.

The crude product, which contained some combined,

hydrogen chloride, is dissolved in methylene chloride (250 ml.), and then methanol (500 ml.) plus triethylamine (5.0 ml.) is added portionwise to the boiling solution until all of the methylene chloride has been removed. The resulting mixture gives 19.61 g. (87% yield) of very deep red crystals, M.P. 242-243 dec., which contains only minor impurities by thinlayer chromatographic analysis and is sufiiciently pure for the next step.

A pure sample is obtained by chromatography on activated magnesium silicate (50-100'mesh) using 15% ether-benzene as eluent. Crystallization of the product from methylene chloride-methanol, as above, gives material of MP. 243 -244 dec.

EXAMPLE 2 I-(p-nitrophenylazo -3-methoxyestra-1,3,5 (10) trien-1 7-0ne To a stirred solution of 17.94 g. (0.04 mole) of 1- (p-nitrophenylazo) -3 -methoxy 4-aminoestra-1,3,5 10) trien-17-one (melting point 242243 C. dec.) in 400 m1. of glacial acetic acid and 100 ml. of 30% (w./w.) aqueous sulfuric acid at 0 C. is added a solution of 3.04 g. (0.044 mole) of sodium nitrite in. 30 ml. of water below the surface of the liquid. Stirring is continued at 0 C. for 15 minutes, then 450 ml. of 50% aqueous hypophosphorous acid is added and the mixture stirred overnight (16 hours) at room temperature. The

precipitate is filtered, washed thoroughly with water and dried to. give 16.4 g. (95% yield) of crude red-brown product, melting point 203 205 C. Crystallization from methylene chloride-methanol gives 15.31 g. (89% yield) of red crystals, melting point 216219 C. which contains only trace impurities by thin layer chromatographic analysis and is pure enough for the next step. Chromatography of a sample on activated magnesium silicate (60-100 mesh), using ether-benzene as eluent, followed by crystallization of .the product from methylene chloride-methanol gives an analytical sample, melting point 223 -224 C.

EXAMPLE3 1-amino-3-methoxyestra-1,3,5 -trien-17-0ne A solution of 13.0 g. (0.03 mole) of 1-p-nitrophenylazo) 3 methoxy 1,3,5(10) estratrien 17 one (melting point 215*217 C. )in 100 ml. of methylene chloride. is added to a stirred mixture of 30 g. of zinc dust in 300 ml. of glacial acetic acid over approximately .10 minutes. The initial temperature of 23 soon rises to 40-45 C. and is maintained in this range during the reaction by occasional cooling in a water bath. An additional 30 g. of zinc dust is added after half of the steroid is fed in. The mixture is stirred for a further 10 minutes then filtered and the residue of zinc washed on the filter with acetic acid. The filtrate is concentrated under reduced pressure to approximately 125 ml., diluted with 500 ml. of water and extracted with chloroform. The extract is washed with two portions of water, dried over magnesium sulfate, concentrated to a .volume of 100 ml. and filtered through a bed of magnesium silicate (60 g.) using 300 ml. of chloroform wash. The filtrate is evaporated under reduced pressure to a small volume and crystallized from methanol to give 5.58 g. of grey solid, melting point 198-208 C. which contains a trace of p-phenylenediamine impurity. An 7 additional 1.0 g. of product, melting point 209 -213 C. is obtained by chromatography of the filtrate on activated magnesium silicate (60-100 .mesh) using 20% ethyl aicetate-mhexane as eluent, followed by crystallization of the product from methylene chloride-ether. The total vyield of product is therefore 6.58 g. (73%).

Analytically pure material is obtained by chromatography of a sample on activated magnesium silicate as above, followed by crystallization from methanol to give a white crystalline product, melting point 213-214 C.

EXAMPLE 4 1-amirw-3-m'eth0xyestra-1,3,5 (1 0) -trien-1 75-01 Sodium borohydride (1.9 g.) is added to a suspension of 1 amino 3 methoxyestra 1,3,5(10) trien 17- one (3.0 g.) in methanol (200 ml.). The resulting mixture is stirred for 30 minutes at room temperature, acidified with acetic acid and evaporated. The residue is partitioned between ether and water, and the ether phase is dried over anhydrous sodium sulfate and evaporated to give the product of the example.

' amino-3-methoxyestra-1,3,5(10)-trien-17 one (melting point 198208 C.) in 200ml. of acetic acid and 139.8 ml. (0.139 equivalent) of 1.0 N sulfuric acid at 20 C. (Dry Ice acetone bath) is added a solution of 3.55 g. (0.051 mole), of sodium nitrite in 100 ml. of water below the surface of the liquid. The solution is stirred for 2 minutes then a solution of 12.1 (0.186 mole) of sodium :azide in 50 ml. of water is added as quickly as the resulting vigorous evolution of nitrogen would allow. The orange colored mixture is stirred at -10 to +5 C. for 30 minutes then extracted with chloroform and the extract washed with water, saturated sodium bicarbonate and finally with water. After drying over magnesium sulfate, the solution is evaporated under reduced pressure to an oil which crystallizes to a dark.

brown solid on cooling. The total crude product is chromatographed in 500 g. of activated magnesium silicate (60-100 mesh) using 7.5% ethyl acetate-n-hexane as eluent. The yield of cream colored solid is 12.1 g. melting point 144-148 C. A single crystallization of a sample from methylene chloride-methanol give-s analytical material with melting point 147 -149 C.

EXAMPLE 6 1-azid0-3-methoxy-1,3,5(10) -estratrien-17;3-0l

Following the procedure of Example 5 1-amino-3- methoxyestra-1,3,5(10)-trien-17/3-ol isconverted into the product of the example.

EXAMPLE -7 1,11-imin0-3-meth0xyestra-1,3,5(1 0) -trienl 7-0ne of crude product which is chromatographed on magne-.

sium silicate (600 g. of 60-100 mesh). Elution with 15% and finally 20% ethyl acetate-petroleum ether (30-75) gives 7.2 g. of product which still contains a less polar impurity. This is removed by dissolving the product in 250 ml. of 0.74 N hydrochloric acid and extracting the solution with ether. The product is recovered by making the aqueous phase basic with excess 5.0 N sodium hydroxide and extracting the precipitate with methylene chloride. Evaporation of the methylene cloride exract under reduced pressure gives an oil which crystallized on trituation with ether giving 6.7 g. of cream colored product, melting point 192-l98 C. Crystallization 5 from methanol gives 5.94 g. (65% yield) of white product, melting point 199 20I C. which is pure by thin layer chromatographic analysis. additional crystallization of a sample from methanol did not alter the melt- EX AM PLE 9 1,11-irhiho 3-inih0xystrd-13,5(10)-tfin-17-one V hydrochloride I Treatment of 1,11 in1ino-3-methoxye'stra-1,3,5(10)- trien-17-onein a mixture of ether and chloroform with hydrogen chloride gives the product of the example.

EXA M PLE 1o 1,11-imin-3-hydroxyestra-1,3,5 (10) -trien-1 7 -one Heating of 1,1l-imino-3-mthoxyestra-l,3,5(10)-trienl7-one (Example in pyridine hydrochloride at a temperature just below the boihng point yields the product of the example.

EXAMPLE 11 1,11-imino-3-hydr0xyestra-1,3,5 -trien-1 713-01 Heating of 1,11-imino-3-methoxyestra-1,3,5 10) -trien-, 175-01 in pyridine hydrochloride at a temperature just below the boiling point provides the product of the example.

EXAMPLE 12 1,11-pr0pi0nylimino-3-methoxyestra-1,3,5 (10) -trien- Treatment of 1,11 imino-3-methoxyestra-1,3,5(l0)- trien-17-one in pyridine with propionic anhydride on the steam bath for 1 hour followed by standing overnight at room temperature gives the product of the example.

EXAMPLE 1 3 1,11-benzoylimino-3-methoxyestra-1,3,5 (10) -trien-1 7-one Substitution of benzoyl chloride for propionic anhydride in Example 12 gives the product of the example.

EXAMPLE 14 1,11- (3,4',5-trimethoxybenzoyl) imino-.i-methoxyesrra- 1,3,5 (10) -trien-17-0ne 1,11-imin0-3-hydr0xyestra-1,3,5 (1 O),-trien-1 7-one hydrochloride Treatment of 1,11 imino 3 hydroxyestra-1 ,3,5(l0)- trien-17-one in a mixture of ether and chloroform with hydrogen chloride gives the desired hydrochloride.

EXAMPLE 16 1 .11imino-3-hydroxyestra-1,3,5 (10) -trien-1 7 -one hydrosulfate Treatment of 1,11 imino 3 hydroxyest-ra-1,3,5(10)- V trien-l7-one in a mixture of ether and methanol with sulfuric acid gives the hydrosulfate salt.

6 EXAMPLE 17 1, l1-acetylimino-3-methoxyestra-1,3,5 (10) -trien-17 -0ne 7 To a solution of 0.5 g. of 1,11-imino-3-methoxyestra- 1,3,5 (10)-trien-l7-one in 10 ml. of pyridine is added 10 ml. of acetic anhydride. The mixture is heated on a steam bath for 1 hour and then allowed to stand at room temperature overnight. The excess acetic anhydride is decomposed with methanol and the solution is evaporated. The residue is partitioned between ether-methylene chloride (2: 1) and 5% sodium carbonate solution. The organic phase is dried and evaporated to give the product of the example.

EXAMPLE 18 1,11 N-methylimin(1-3-meth0xye.stra 1,3,5 (10) -'trien-17-0'ne A solution of 1,1l-imino-3methoxyestra-1,3,5(l0)- trien-:17-one in 20% dimethylttormamide-benzene is treated with sodium hydride and methyl iodide. mixture is heated at 40 C. for 1 hour, cooled and extracted with benzene. The extract is washed with water and evaporated to give the N-methyl derivative, the product of the example.

EXAMPLE l9 1,11-imin0-17a-ethynyleSt/'a-1,3,5 (10)- mane-3,1 713-211'01 A solution of potassiurn-acetylide in liquid ammonia is prepared by bubbling acetylene into a solution containing 3 g. of potassium in 150 ml. of liquid ammonia until the blue color disappears. To it is added dropwise a solution of 1.3 g. of 1,.11-imino-3 hydroxyestra- 1,3,5,(10)-trien-17-one in 75 'ml. of ether. After 2 hours the cooling bath is removed and the mixture is allowed to warm to room temperature and stored overnight. Dilute sulfuric acid (8%, ml.) is added dropwise to the mixture and it is extracted with ether. The extract is washed with saturated sodium chloride solution, dried and evaporated to give the product of the example.

EXAMPLE 2 0 1,1 I-imino-l 7u-vinylestra-1,3,5 (10) triene-3,17B-diol 1,11-imin'0-17ot-methylestra-1,3,5(10) triene-3,1 7fl-di0l To a freshly prepared solution of methyl magnesium iodide (4.2 ml. of methyl iodide and 1.6 g. of magnesium turnings) in ether (30 ml.) is added a solution of 1,11- imino-3-hydroxyestra-1,3,5(10)-trien-17-one (0.9 g.) in ether (25 ml.). The resulting mixture is heated to reflux for 6 hours, cooled and quenched-by the dropwise addition of excess ammonium chloride solution. The ether phase is separated and the aqueous phase is extracted several times with methylene chloride. The combined extracts are washed with water, dried and evaporated to give the product of the example.

EXAMPLE 22 1,11-N-methylimino-3-meth0xyestra-1,3,5 (10) trien-17-one methiodide A solution of 1,ll-N-methylimino-B-methoxyestra- 1,3,5 (10)-trien-17-one in either-methanol is treated with methyl iodide to give the methiodide.

The.

7 We claim: 1. A compound of the formula:

wherein R is selected from the group consisting of hydrogen and lower alkyl; R is selected from the group consisting of hydrogen, lower alkyl, lower alkanoyl, mononuclear aroyl, and lower alkoxy mono-nuclear aroyl; X is selected from the group consisting of C=O;

on loweralkyl (MIL-H, (Eon andacid addition and quaternary ammonium salts thereof.

2. the compound 1,1'1 imino 3 methoxyestra- 1,3,5 l0) -t rien-17-one.

3. The compound 1,11 imino 3 hydroxyestra- 1,3,5 -trie'n-17-one.

4. The compound 1,11 imino 3 methoxyest-ra- 1,3,5(10)-trien-17-one hydrochloride.

5. The compound 1,11 imino 3 hydroxyestra- 1,3,5 (10) -t'rien-17-one hydrochloride.

6.The compound 1,11' imino 3 hydroxyestra 1,3,5(l0)-trien-l7-one hydrosulfate. r

7. The compound 1,11 acetylimino 3 methoxyestra-l,3,5(10)-trien-17-0ne.

8. The compound 1,11 imino 3 methoxyestral,3,5(10)-trien-17fi-ol.

9. The compound 1,11 propionylimino 3 methoxyestra-l ,3,5 10) -t.rien-17-one.

10. The compound 1,11 benzoylimino 3 methoxyestra-1,3,5(10)-trien-l7-one. 11. The compound 1,1l-(3',4,5'-trirnethoxybenzoyl)- imino-B-methoxyestna-1,3,5(10)etrieu-17-one.

12. The compound 1,11 imino 17a ethyny1estra- 1,3,5(10)-triene-3,17fl-diol.

13. The compound 1,11 imino 17c: vinylestra- 1,3,5 10)-triene-3-17,B-diol.

14. The compound 1,11 imino 17m methylestra- 1,3 ,5-(l0)-triene-3,l7fi-diol.

15. The compound 1,11 N methylimino 3 methoxyestra-1,3,5 10) -trien-17-one.

16. The compound 1,1 1 N methylimino 3- methoxyestra-1,3,5 10) -trien-1 7-'one methiodide.

17. The compound 1 azido 3 methoxyestral,3,5(l0)-trien-17-one.

18. The compound 1 azido 3 methoxyestra- 1,3,5(10)-trien-17}3-0l.

No references cited.

LEWIS GO'ITS, Primary Examiner.

ELB'ERT L. ROBERTS, Examiner. 

1. A COMPOUND OF THE FORMULA: 